摘要
目的 :研究药物敏感虫株伯氏疟原虫K173株及其在本芴醇压力下产生的本芴醇抗性株之间基因表达的差异状况 ,克隆抗性相关基因。方法 :采用mRNA差异显示技术寻找相关基因 ,并用Northern杂交证实。结果 :采用 2 4种引物组合进行mRNA差异显示 ,克隆到 30余个差异表达基因片段 ,对其中 8个进行了Northern杂交鉴定及测序。结果表明 ,与敏感株相比 ,克隆CB52在本芴醇抗性株中表达明显增多 ,而在氯喹抗性株中的表达有所减少 ,说明克隆CB52确与本芴醇抗性相关 ,序列同源性检索表明CB52序列与恶性疟原虫环亲蛋白基因有微弱同源性。向GenBank登录 8条新基因序列。结论 :克隆CB52基因表达水平的改变与伯氏疟原虫本芴醇抗性的产生相关 。
Objective:To isolate genes related to benflumetol resistance in the rodent malaria P.berghei K173 strain.Methods:The mRNA differential display technique was used to identify differentially expressed genes between drug sensitive P.berghei K173 strain and benflumetol resistant strain selected under benflumetol pressure.Northern blot was performed in order to determine the genes actually derived from maternal strains.Results:PCR reactions were prepared respectively with twenty four pairs of primers in each of the strains.Thirty three differentially displayed genes fragments were cloned and eight of them were sequenced. The eight sequences were novel ESTs and were submitted to GenBank. Homology searching revealed that clone CB52 was slightly homologous to P.falciparum cyclophilin gene. Hybridization confirmed that the gene fragment CB52 was upregulated in the benflumetol resistant parasite strain, while downregulated in the chloroquine resistant strain.Conclusions:CB52 fragment was involved in the benflumetol resistance in P.berghei and the genetic mechanisms of drug resistance to benflumetol and chloroquine were different. [
出处
《军事医学科学院院刊》
CSCD
北大核心
2001年第2期107-110,共4页
Bulletin of the Academy of Military Medical Sciences
关键词
伯氏疟原虫
本芴醇
氯喹
基因表达
抗药性
Plasmodium berghei
benflumetol
chloroquine
gene expression
drug resistance
